Temporal traumatic brain injury biomarkers and methods of use thereof

Abstract

A unique pipeline is employed for biomarker discovery that entailed domain antibody phage display, next generation sequencing analysis, and nanotechnology strategies to generate antibody mimetics are disclosed. Also disclosed are the temporal biomarkers of traumatic brain injury and their methods of use. In some embodiments, the temporal biomarkers are synthetic peptides comprising the HCDR3 sequences identified using the disclosed pipeline. In some aspects, the synthetic peptides have less than 30 amino acid residues and comprise a biotin scaffold that is linked to the HCDR3 sequences.

We claim: 
     
       1. A peptide having less than 30 amino acid residues comprising a recognition sequence selected from the group consisting of: TAERDARTFQY (SEQ ID NO. 1), SLYGSSRHTAPISF (SEQ ID NO. 2), TDLAVAHPVRY (SEQ ID NO. 3), AAPSWNNHVSY (SEQ ID NO. 4), RLVRESSQEHTLSS (SEQ ID NO. 5), TDCQETPYELKS (SEQ ID NO. 6), TGHEGENEMAS (SEQ ID NO. 7), GPLDGKEEELRF (SEQ ID NO. 8), or GGDTFRDASQSMHF (SEQ ID NO. 9). 
     
     
       2. An imaging composition, the composition comprising at least one of the peptides of  claim 1 . 
     
     
       3. The peptide of  claim 1 , wherein the peptide further comprises an N-terminal cysteine and a C-terminal cysteine. 
     
     
       4. The peptide of  claim 1 , wherein peptide is biotinylated. 
     
     
       5. The peptide of  claim 4 , wherein the peptide comprises a biotin scaffold having X1-X2-(X3-X4)-Gly-Ser-DLys-Ser-Gly-Ser(Biotin)-Gly-PropargylGly (SEQ ID NO. 10), wherein:
 X1 and X3 each is any amino acid, 
 X2 and X4 each is any amino acid or absent, and 
 X1 and X3 of the biotin scaffold each is bonded to the terminal residues of recognition sequence. 
 
     
     
       6. The peptide of  claim 5 , wherein X1 is E and X2 is K in SEQ ID NO. 10. 
     
     
       7. The peptide of  claim 5 , wherein X3 is E and X4 is absent in SEQ ID NO. 10. 
     
     
       8. The peptide of  claim 5 , wherein X1 is E, X2 is K, X3 is E, and X4 is absent in SEQ ID NO. 10. 
     
     
       9. A method of identifying a site of brain injury in a subject comprising
 administering to the subject a targeting peptide comprising a recognition sequence selected from the group consisting of TAERDARTFQY (SEQ ID NO. 1), SLYGSSRHTAPISF (SEQ ID NO. 2), TDLAVAHPVRY (SEQ ID NO. 3), AAPSWNNHVSY (SEQ ID NO. 4), RLVRESSQEHTLSS (SEQ ID NO. 5), and TDCQETPYELKS (SEQ ID NO. 6), wherein the targeting peptide has less than 30 amino acid residues; 
 detecting the targeting peptide at a site in the brain of a subject; and 
 identifying the site of brain injury in the subject at the site the targeting peptide is detected in the brain of the subject. 
 
     
     
       10. The method of  claim 9 , wherein the targeting peptide is administered to the subject via an intravenous injection or an intraspinal injection. 
     
     
       11. A method of identifying a site of brain injury, the method comprising:
 providing a brain tissue sample; 
 providing a targeting peptide having less than 30 amino acid residues and comprising a recognition sequence selected from the group consisting of: TAERDARTFQY (SEQ ID NO. 1), SLYGSSRHTAPISF (SEQ ID NO. 2), TDLAVAHPVRY (SEQ ID NO. 3), AAPSWNNHVSY (SEQ ID NO. 4), RLVRESSQEHTLSS (SEQ ID NO. 5), and TDCQETPYELKS (SEQ ID NO. 6); 
 bringing the targeting peptide into contact with the brain tissue sample; 
 illuminating the brain tissue sample; and 
 detecting light from the brain tissue sample in response to the illuminating light, 
 wherein the location of detected light from the brain tissue sample is the site of brain injury. 
 
     
     
       12. The method of  claim 11 , wherein the targeting peptide comprises a recognition sequence selected from TAERDARTFQY (SEQ ID NO. 1) and SLYGSSRHTAPISF (SEQ ID NO. 2), the location of detected light from the brain tissue sample is the site of acute brain injury. 
     
     
       13. The method of  claim 9 , wherein the targeting peptide comprises a recognition sequence of SLYGSSRHTAPISF (SEQ ID NO. 2), the site of injury identified is a site of acute brain injury or received trauma to the brain within 24 hours. 
     
     
       14. The method of  claim 13 , further comprising administering to the subject a second targeting peptide, wherein the second targeting peptide has less than 30 amino acids residues and comprises a recognition sequence selected from the group consisting of TAERDARTFQY (SEQ ID NO. 1), TDLAVAHPVRY (SEQ ID NO. 3), AAPSWNNHVSY (SEQ ID NO. 4), RLVRESSQEHTLSS (SEQ ID NO. 5), and TDCQETPYELKS (SEQ ID NO. 6). 
     
     
       15. The method of  claim 11 , further comprising:
 providing at least one second targeting peptide, wherein the second targeting peptide has less than 30 amino acids residues, is different than the targeting peptide, and comprises a recognition sequence selected from the group consisting of TAERDARTFQY (SEQ ID NO. 1), SLYGSSRHTAPISF (SEQ ID NO. 2), TDLAVAHPVRY (SEQ ID NO. 3), AAPSWNNHVSY (SEQ ID NO. 4), RLVRESSQEHTLSS (SEQ ID NO. 5), and TDCQETPYELKS (SEQ ID NO. 6); and 
 bringing the second targeting peptide into contact with the brain tissue sample. 
 
     
     
       16. The method of  claim 15 , wherein the targeting peptide comprises a recognition sequence of TAERDARTFQY (SEQ ID NO. 1) and at least one second targeting peptide comprises a recognition sequence of SLYGSSRHTAPISF (SEQ ID NO. 2), the location of detected light from the brain tissue sample is a site of acute brain injury. 
     
     
       17. The method of  claim 15 , wherein the at least one second targeting peptide comprises a recognition sequence of selected from TDLAVAHPVRY (SEQ ID NO. 3) and AAPSWNNHVSY (SEQ ID NO. 4), the location of detected light from the brain tissue sample includes a site of subacute brain injury. 
     
     
       18. The method of  claim 15 , wherein the at least one second targeting peptide comprises a recognition sequence of selected from RLVRESSQEHTLSS (SEQ ID NO. 5) and TDCQETPYELKS (SEQ ID NO. 6), the location of detected light from the brain tissue sample includes a site of chronic brain injury. 
     
     
       19. The method of  claim 15 , wherein the target peptide and the at least one second targeting peptide comprise a biotin scaffold having the sequence: 
       
         
           
                 
               
                   (SEQ ID NO. 10) 
                 
                   X1-X2-(X3-X4-)-G-S-K-S-E-K(Biotin)-G-PropargylG 
                 
             
                
                
               
            
           
         
         wherein:
 X1 and X3 each is any amino acid, and 
 X2 and X4 each is any amino acid or absent, and 
 
         the target peptide and the at least one second targeting peptide further comprise a cysteine residue at the C-terminus and N-terminus, wherein X1 and X3 of the biotin scaffold each forms a bond with the N-terminal cysteine and the C-terminal cysteine of the target peptide and the at least one second targeting peptide. 
       
     
     
       20. The composition of  claim 2 , wherein the peptide comprises a recognition sequence selected from TAERDARTFQY (SEQ ID NO. 1) and SLYGSSRHTAPISF (SEQ ID NO. 2), the peptide identifies the subject received trauma to the brain within 24 hours.